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When cell differentiation and matrix mineralization were evaluated after days 7 and 28, the MWCNT-COOH group showed increased cell differentiation and matrix mineralization, when compared to the MWCNT group.
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In vitro mineralization was evaluated based on histochemical staining and quantification of the hydroxyapatite content of the extracellular bone matrix.
The osteogenic differentiation was assessed by alkaline phosphatase (ALP) activities/staining and the mineralization was evaluated by Alizarin red S staining.
Extracellular matrix mineralization was evaluated using Alizarin Red Staining.
Mineralization was evaluated by Alizarin red staining.
In vitro mineralization was evaluated by alizarin red staining.
The long-term mineralization was evaluated by calcium content deposited by the cells and a Von Kossa staining after 30 days of culture.
Cell proliferation, apoptosis, alkaline phosphatase (ALP) activity, and mineralization was evaluated by methylthiazoletetrazolium (MTT) and bromodeoxyuridine (BrdU) assay, flow cytometry, ALP viability kit and von Kossa staining, respectively.
Mineralization was evaluated by nodule formation on days 7, 14, and 21 after differentiation with the osteogenic media (α-MEM with 10 mM β-glycerophosphate (Sigma-Aldrich, St . Louis MO, USA) and 0.2 mM ascorbic acid (Sigma-Aldrich, St . Louis MO, USA)).
Mineralization was evaluated by the von Kossa staining of cells after 30 days of culture in the presence or not of 10 ng/ml bone morphogenic protein-2 (BMP-2).
Specifically, hMSC's adhesion, proliferation, mineralization, and differentiation were evaluated.
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